Download National Forum on State an d Challenges of UTILISATION OF

DAVID A. MBAH, PhD
CAMEROON ACADEMY OF SCIENCES

« Science and other scholarly endeavours can and must
inform public policy but the public is the ultimate decision
maker about which problems are most pressing, which risks
are most worth taking, and which technologies are most
desirable given all the available alternatives »
(Beth Elpern Barrows, 2002)

It is hoped that at the end of the presentation,
participants will be more aware of:
• Genetic modification
• Natural and artificial forces driving genetic
modification
• Difference between conventional and modern
genetic modification
• Intended and untended effects/risks
• Management/control of unintended effects/risks
Outline:

 Definition of terms
 Process of GM of animals :
 Developmemnt of process of modern GM
 Application of modern GM in animals
Genetic modification : Conventional and Modern;
- Natural forces
- Artificial forces
- Target traits for Cameroon
- Unintended effects
- Control/management of unintended effects
- Summary
Definition of terms
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Gene: basic hereditary unit (made up of DeoxyriboNudeic Acid-DNA) which
determines protein structure or Ribo Nucleic Acid-RNA). It is located at a
point (locus) on a chromosone. Each member of a pair of genes at the locus
is called an allele.
Genotype: genetic identity of an individual (e.g.AA, Aa, aa where A and a
are alleles)
Phenotype: outward manifestation of genetic identiy, often in interaction
with environment
Genetic engineering: changing genetic constitution by
introduction/elimination of gene(s): Genetic Surgery
Transgene: a gene construct introduced from another species into an
organism by human intervention using modern genetic modification
measures
Genetic modification: change in gene and genotypic frequencies among
individuals of each generation
Comparator: product that is compared to another product (e.g. G.M
organism versus non-GM organisl/GM food versus non-GM food).
Substantial equivalence: « as safe as its conventional counterpart »

The procedure for producing transgenic microbes (Cohen,
1975) was adapted for transgenic animals by Smith (1996).
• Identification/construction of foreign gene
• Microinjection of identified DNA (gene) and pronucleus
of a fertilized egg
• Implantation of resulting recombinant(chimera) eggs
(cells) into surrogate
• Development of embryo to term
• Proving that foreign DNA has been stably and heritably
incorporated in the DNA of at least some to the newborn
offspring
• Demonstrating that the gene expresses itself in the new
environment (recombinant)
 Using this procedure: rat gene (for growth
homone) was inserted in mouse genome.
 It expressed itself producing progeny that were
much larger than parents (Nicholl, 1994) => first
Transgenic animals
Since then: Table 1 and Brophy et al(2003) on
cloned transgenic dairy cows. Cloning GM cattle
does not appear to negatively modify meat and milk
compositions(Tian et al, 2005; van Berkel et al, 2007) =
substantial equivalence(genetic and breed
comparators)
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

• Concerns/Risks:
=> conventional GM
=> modern GM
• Controlling concerns/risks
Genetic Modification (1)

Normal Situation:
• Genetic structure of population of animals determined by proportions of
different genotypes in the population (Falconer, 1989) (e.g. for locus « A »,
the number of AA, Aa, aa individuals in a population of 100?)
• Proportions in current generation determine proportions in next
generation
• For locus A in entire population, assume:
- frequency of A = P
- frequency of a = q
-p+q=1
and that for next generation, gametes produced are:
AA => A gametes (1 type)
Aa => A and a gametes (2types
aa => a gmetes (1type)
Genetic Modification (2)
The result at single locus (A) is

Male/Female
P(A)
q(a)
p(A)
P²(AA)
pq(Aa)
q(a)
qp(aA)
q²(aa)
Ratio is p²(AA): 2pq(Aa): q²(aa)
• When: p & q remain unchanged
•
There is random mating
Genetic modification (3)

 Disturbed Situation: forces acting on normal situation
I Natural forces
1. Mutation: heritable change in gametes from one allele to another
U
A <=> a
V
If U=V, mutation has no effect
If U≠V, mutation is directional and effective, Hence, p and q be
modified (increased or decreased)(e.g. Sickle Cell gene and Albino
gene)
• The genetic structure of the next generation will depend not
only on the gene frequencies in the preceding generation but
also on the mutation rate
Genetic Modification (4)

2. Survival forces
* Fitness – some alleles affect ability to survive to
reproductive age
- some alleles affect ability to produce viable
offspring
- some genotypes die early or are handicapped
in one way or another relative to mating
chances
Result: « fitter » individuals (genotypes) determine the genotypes
and gene frequency for the next generation. Such a generation has
fewer or no « defective » genes. It is therefore genetically different
from the preceding generation.
Genetic Modification (5)

3. Migration (Introgression)
A population of animals may be composed of
« natives » and « immigrants ». The « immigrants » may
modify the genetic structure of the population by
increasing a given gene frequency.
For given locus (Fatconer, and Mackay, 1996)
qo = gene frequency among natives
qm = gene frequency among immigrants
q1 = gene frequency among mixed population
q1 = mqm + (1-m)qo = m(qm-qo) + qo
Genetic modification (6)

 Where: m = proportion of new
immigrants/generation
∆q1 due to one generation of new
immigrants:
qm = q1 – qo
= m(qm- qo)
Thus the rate of change in gene frequency in a
population undergoing immigration
(introgression) depends on:
a) immigration rate (m)
b) difference in gene frequency between
« immigrants (qm) and « natives » (qo).
Genetic modification (7)

i Animal breeding => Sexual reproduction
(a) selection is based on defined criteria to
determine individuals for mating to produce
individuals with given traits in the next generation
(b) Crossbreeding to combine breed
characteristics/differences in chosen traits for desired
results.
(a) + (b) are based on the « breeding value » (i.e. value
associated with genes carried by individuals and
transmitted to their progeny/ offspring.
Genetic modification (8)

 Measure of breeding value: « average effect » (Falconer &
Mackay, 1996: 112-114)
The average effect may be assigned to
(a) a gene in the population or
(b) the difference between one gene and another of an
alleleic pair.
The « average effect of a gene » then is the « mean deviation
from the population mean of individuals which received the
gene from one parent, the gene from the other parent having
come randomly from the population » (e.g. 20 calves
receiving a gene A, from 1 bull mating 40 cows which done
the other allele, a).
Genetic modification (9)

This is easier seen as the « average effect of gene
substitution » at one locus of 2 alleles (e.g. changing
« A » to « Aa -> aa) in a population
The average effect of gene substitution depends on the
gene and the population. It is high when gene
frequency is high and low when gene frequency is low.
Genetic modification (10: 1 - 3)

Results from Wakwa IRAD Centre show how during a
17 year period selection modified the genetic structure
of Gudali beef cattle population (Ebangi et al, 2002)
with long generation intervals (7-8 years) (Tawah et al,
1993)
Fig 1: Direct and maternal gentic trends for yearling weight in Gudali beef
Cattle. EBV = estimated breeding value


Comment: Total Direct Gain: +5.5 kg EBV
Total Maternal Gain: -2.5 kg EBV
Total Genetic Trend: + 3.0 kg EBV
(resulting from negative genetic
correlation of -0.81)
HOW MANY GENES WERE INVOLVED ?
Crossbreeding (1): NO. OF GENES ?

Gudali bull (TDCS)
Holstein(62.5%)Gudali(37.5%) bull
(TDCS)
Crossbreeding (2): NO. OF GENES ?

Brahman(50%)-Gudali
(50%)bull (TDCS)
Simmental(50%)-Gudali
(50%)bull (TDCS)

Genetic modification(11)
Animal Production problems needing GM (CAS, 2014;
Vermaak et al, 2015; The Roslin Institute, 2015):
• African Swine Fever(Swine production)
• Foot and Mouth Disease(Cattle production)
• Coccidiosis(Poultry production)
• Bird Flu(Poultry production, Human health)
• Trypanosomiasis(Cattle production)
• African Horse Sickness(Horse production)

Genetic modification (12)
Unintended effects of selection/crossbreeding (1)
Success in selected trait may lead to undersirable
consequences:
* negative genetic correlation as in EBV above
* increased size => increased pressure on the
environment
* increase in fitness (reproductive) => need for
more space => pressure on the environment and other
species
* genetic erosion.

Genetic modification(13)
Management/Handling of Unintended Effects
of selection/crossbreeding (2)
- involving more than one trait in selection
- determinning carrying capacities for
given pasture lands, etc
- off-take rate to allow « ecologically
sound » stocking rate
- conservation (in situ and/or ex situ) of
displaced genes (breeds)
Genetic modification (14)

ii. Transgenesis/Genetic Engineering
Transgene (tg) (Braig and Yan, 2002): Assume that
qtg = Atg ; e.g. 100 = 1/20 =0.05
2N
(2) (1000)
Where n = 100 = number of alleles of tg in the population
N = 1000 = population size of the diploid population
If q increases, transgenic individuals (GMOs) have tg
advantage over their wild types.
Genetic Modification (15)

The transgene may:
1. Introduce a novel/new trait in the population
where the trait did not exist (i.e. q = 0)
2. Increase the intensity (where trait exists i.e. qtg > 0)
/\ qtg = qtg1 - qtg0
Where qtgo > 0 in the population,
When the initial qtg is « above the critical threshold »
density, the transgene can be spread and fixed within
60-100 generations. HOW MANY GENES INVOLVED?

1. Genetic modification(16: 1 – 2)
2. Environmental effects: Transgenic animal
entering the environment through release or
escape
Transgene could spread through:
* vertical transmission (reproduction with
wild relatives)
* horizontal transmission by vectors.


2. Genetic modification(17)
3. Food safety (products from beef/dairy, cattle,
sheep/goats, poultry and eggs, pigs, rabbits,
fishes, etc): New/introduced genes could lead to
new proteins which may have the following
effects
* allergenicity
* bioactivity (of molecules enhancing
growth, etc)
* toxity
Genetic modification (18)
Pharmaceutical production:
Table 3: Transgenic Animals as Producers of Pharmaceuticals
Species
Chicken
Rabbit
Goat
Sheep
Cow
Pharmaceutical Product

Monoclonal antibodies

Lysozyme

Growth hormone

Insulin

Human Serum albumin

Calcitonin

Superoxide dismutase

Erythropoietin

Growth hormone

IL-2

X-glucosidase

Ari
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




Antithrombin III
Tissue plasmogen activator
Monoclonal antibodies
X-1-Antitrypsin
Growth hormone







x-1-Antitrypsin
Factor VIII
Factor IX
Fibrinogen
Human serum albumin
Lactoferrin
x- Lactalbumin
Source: NAS.(2002 ), van Berkel et al(2002)

Genetic modification(19)
3. Animal health: Ruminants (cattle, sheep, goats) produced
through nuclear transfer methods containing transgenes (or
not) could lead to;
* higher birth weights
* longer generation lengths than for calves/lambs
from artificial insemination
Comment: Efficiency of methods: Extremely inefficient (0 – 4% in
cattle, sheep, goats, pigs).
* Mortality: 80-90% during early development
* Many survivors show improper expression of
inserted gene.
* Many survivors show abnormalities: anatomical,
physiological, behavioral
Genetic modification(20)
Management/Control of the Risks (Cameroon
Biosafety Manual, 2004: 30-31; MINEPDED: Cam Bio
Proj: Ongoing): Modern GM
1.

Risk assessment: Case by case
* Concern about GM activity identified
* Potential harm evaluated
* Likelihood and consequence of harm
determined
* risk management procedure(s) investigated
* acceptance of the risk evaluated
* activity is either:
a) approved with or without risk
management, or
b) refused until more information
is available.
2.
3.

Genetic modification(21)
Risk Management:
* differs from animal to animal, gene to gene
* focuses on containing the GM animal and novel
gene(s) within the activity area, minimising
accidental release, ensuring removal of the GM animal
from the environment after testing.
* applicant proposes risk management methods (to
minimize likelihood of real harm)
* biosafety review team assess the proposed
measures
* biosafety review team approves the measures or
recommends modification
* regulators append recommended RM measures
as
conditions to authorization

Summary

Genetic modification (process whereby gene frequencies and genetypic
frequencies are modified among individuals of each generation) of animals is
driven by natural and artificial forces.
Natural forces include mutation, fitness and migration/introgression.
Artificial forces include selection, crossbreeding and transgenesis/genetic
engineering.
Genetic modification driven by natural forces is essentially adaptive while
modification driven by artificial forces is controlled by human intervention
aimed at meeting food, health and other needs.
Conventional genetic modification under sexual reproduction within species
produces both beneficial and negative effects. Modern genetic modification
– interspecific exchange of genes using genetic engineering – has beneficial
and negative effects as well at varying degrees depending on species
involved. Control/management systems/mechanisms are developed and
applied to enable societal benefits while minimizing/preventing negative
effects of conventional and modern genetic modification. Targeted analysis
of selected nutrients in animal products is made on a case-by-case basis to
test substantial equivalence of any compositional changes resulting from
genetic modification, Unique identifiers are established to track GM animals
and animal products in the food chain,
References

1.
Barrows B.E. (eds): Genetically engineered Organisms: Assessing Environmental and
huamn health effects. CRC Press, Washington, D.C. pp 251-314
2.
Beth Elpern Barrows. 2002. In: D.K. Letoumeau, Bath Elpern Bussows (Eds): Genetically
Engineered Organisms: Assessing Environmental and Human Effects
3.
Braig H.R. and Yan G. 2002. The spread of genetic contructs in natural insect populations.
In: Le Tourneau D.K.,
4.
Brophy B., Smolenski, Wheeler T., Wells D., L’huillier, P., Laide G. 2003. Cloned transgeneic
cattle produce milk with higher levels of B-casein and K-casein Nat. Biotechnology 21: 157162
5.
CAS. 2014. Elements for national biotechnology policy framework in Cameroon.
Cameroon Academy of Sciences. www.casciences.com
6.
Cohen S.N. 1975. The Manipulation of genes. In: Freifelder D. (Ed.): Recombinant DNA,
Scientific American, Freeman. Pp. 113-121
7.
Ebangi L.A., Erasmus G.J., Tawah C L., Mbah D.A. 2002. Genetic trends for growth in a
selection experiment involving purebred and two-breed synthetic beef breed in a tropical
environment. Rev. Elev. Med. Vet. Pays trop. 55 (4) 305-212
8.
Falconer D.S., Mackay F.C. 1996. Quantitative Genetics (23-136). Pearson/Preatice Hall

9.
Götz Laible*, Brigid Brophy, Derek Knighton, David N. Wells (2007) Compositional
analysis of dairy products derived from clones and cloned transgenic cattle: pp. 166-167
10.
Mbah D.A. 2006. Genetic Modification: natural selection, artifical selection and genetic
engineering. JCAS 6(1): 19-24
11.
Mbah D.A. 2006. Biotechnology and Animal Production. JCAS 6(1): 29-40
12.
MINEPDED. 2015. Consultancy reports for « Devepment and Institution of a National
Monitoring and Control System for Living Modified Organisms and Invasive Alien
Species »
13.
National Research council (NRC). 2002. Animal Biotechnology: Science-Based Concerns.
Pp. 181. The national Academies Press, Washington, D.C.
14.
National Academy of Sciences (NAS) 2004. Safety of Genetically Engineered Food:
Approaches to assessing unintended health effects. Pp.235
15.
Patrick H.C. van Berkel1*, Mick M. Welling2, Marlieke Geerts1, Harry A. van Veen1, Bep
Ravensbergen3, Mourad Salaheddine1, Ernest K.J. Pauwels2, Frank Pieper1, Jan H.
Nuijens1, and Peter H. Nibbering3: 2002. Large scale production of recombinant human
lactoferrin in the milk of transgenic cows, Nature biotechnology , vol. 20 pp. 484-487
13, Vermaak E., Paterson D. J., Conradie A., Theron J. 2015.
Directed Genetic Modification of African Horse Sickness
Virus By Reverse Genetics. SAJS 111(7/8): 1 – 8
14. The Roslin Institute. 2015. Chickens that don’t Transmit
Bird Flu. http://www.roslin.ed.ac.uk/public-interest/gm chickens. Accessed 12/8.2015
.
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Other References (attention: Scientists, Regulators)
Hilbeck A. and Andow D. A. (Eds). 2004. Environmental Risk
Assessment of Genetically Modified Organismes Vol. 1: A case
study of Bt maize in Kenya. CABI Publishing. Pp. 281
Hilbeck A. Andow D.A. and Fontes ENG. 2006. Environmental
Risk Assessement of Genetically Modified Organisms. Vol. 2:
Methodologies for assessing BT cotton in Brazil. CABI
Publishing.